血浆血结素的测定对自身免疫性溶血性贫血诊断的临床意义

观察３２例自身免疫性溶血性贫血（ＡＩＨＡ）患者血结素含量变化，并与阵发性睡眠性血红蛋白尿（ＰＮＨ）患者的血结素含量进行对比分析，结果显示，ＡＩＨＡ患者血结素含量减低，且与溶血患者的病情相关。提示测定血结素含量对ＡＩＨＡ的诊断与溶血病情的估计有临床意义。

Necrotizing enterocolitis: A multifactorial disease with no cure

Necrotizing enterocolitis is an inflammatory bowel disease of neonates with significant morbidity and mortality in preterm infants. Due to the multifactorial nature o the disease and limitations in disease models, early diagnosis remains challenging and the pathogenesis elusive. Although preterm birth, hypoxic-ischemic events formula feeding, and abnormal bacteria colonization are established risk factors, the role of genetics and vasoactive/inflammatory mediators is unclear Consequently, treatments do not target the specific underlying disease processes and are symptomatic and surgically invasive. Breast-feeding is the most effective preventative measure. Recent advances in the prevention of necrotizing enterocolitis have focused on bioactive nutrients and trophic factors in human milk. Developmen of new disease models including the aspect of prematurity that consistently predisposes neonates to the disease with multiple risk factors will improve our understanding of the pathogenesis and lead to discovery of innovative therapeutics.

Relationship between serum calcium and CA 19-9 levels in colorectal cancer

AIM:To examine the calcium metabolism of colorectal cancer (CRC)in patients with colorectal cancer and control patients. METHODS:Seventy newly diagnosed CRC patients were included.The healthy control group was age and gender matched(n=32).Particular attention was devoted to the relationship between serum calcium of patients,and levels of AFP,CEA,carbohydrate antigen 19-9(CA 19-9)(that could be considered as prognostic factors).Furthermore,the Ca-sensing receptor(CaSR)gene A986S polymorphism was investigated in these patients,as well as the relationship between different CaSR genotypes and the data stated above. RESULTS:A lower level of ionized calcium(also corrected for albumin)was found in the serum of CRC patients with normal 25(OH)vitamin D levels.The ionized calcium concentration was inversely correlated with the serum level of CA.19-9.There was no difference in the distribution of CaSR genotypes,between CRC patients and general population.The genotypes did not correlate with other data examined. CONCLUSION:Based on these results,lower levels of serum calcium might be a pathogenic and prognostic factor in colorectal cancer.

Development of hepatorenal syndrome in bile duct ligated rats

AIM: To evaluate in bile duct ligated rats whether there were progressive alterations of renal function without changes in histopathology. METHODS: Male Wistar rats were submitted to sham-surgery or bile duct ligation (BDL) and divided according to the post-procedure time (2, 4 and 6-wk). To determine renal function parameters, rats wereplaced in metabolic cages and, at the end of the experiment, blood and urine samples were obtained. Histology and hydroxyproline content were analyzed in liver and renal tissue. RESULTS: Rats with 2 wk of BDL increased free water clearance (P = 0.02), reduced urinary osmolality (P = 0.03) and serum creatinine (P = 0.01) in comparison to the sham group. In contrast, rats at 6 wk of BDL showed features of HRS, including signif icant increase in serum creatinine and reductions in creatinine clearance, water excretion and urinary sodium concentration. Rats with 4 wk of BDL exhibited an intermediate stage of renal dysfunction. Progressive hepatic f ibrosis according to post-procedure time was confirmed by histology. The increased levels of liver hydroxyproline contrasted with the absence of structural changes in the kidney, as assessed by histology and unchanged hydroxyproline content in renal tissue. CONCLUSION: Our data show that BDL produced progressive renal dysfunction without structural changes in the kidney, characterizing HRS. The present model will be useful to understand the pathophysiology of HRS.

CONSTRUCTION AND IMMUNOGENICITY OF HUMAN PAPILLOMAVIRUS TYPE 6B L1 RECOMBINANT PLASMID

To construct a DNA vaccine as a prophylactic model to prevent condyloma acuminatum and detect its immu-nogenicity in mice. Methods The major capsid protein (L1) gene of human papillomavirus (HPV) 6b was inserted into an eukaryotic ex-pression plasmid (pcDNA3.1). The recombinant plasmid was transfected into COS-7 cells. Western blot were performed to detect whether L1 protein can be expressed in eukaryotic cells. Eighteen female BALB/c mice were tested for immunoge-nicity study. Results The recombinant plasmid (pcDNA3.1-HPV6bL1) was verified as HPV6b L1 gene by sequencing. Western blot showed specific strip. Anti-L1 protein antibodies could be detected in the mice’s sera inoculated with pcDNA3.1-HPV6bL1. Similarly, IL-4, IL-2, and IFN-γ were increased in the same mice. Conclusion HPV6b L1 recombinant plasmid was constructed successfully which had immunogenicity for BALB/c mice. It provided experimental evidence for the research of DNA vaccine of condyloma acuminata..-

Gallbladder motor function, plasma cholecystokinin and cholecystokinin receptor of gallbladder in cholesterol stone patients

AIM: To study the interactive relationship of gallbladder motor function, plasma cholecystokinin (CCK) and cholecystokinin A receptor (CCK-R) of gallbladder in patients with cholesterol stone disease.METHODS: Gallbladder motility was studied by ultrasonography in 33 patients with gallbladder stone and 10 health subjects as controls. Plasma CCK concentration was measured by radioimmunoassay in fasting status (CCK-f) and in 30 min after lipid test meal (CCK-30).Radioligand method was employed to analyze the amount and activity of CCK-R from 33 gallstone patients having cholecystectomy and 8 persons without gallstone died of severe trauma as controls.RESULTS: The percentage of cholesterol in the gallstone composition was more than 70%. The cholesterol stone type was indicated for the patients with gallbladder stone in this study. Based on the criterion of gallbladder residual fraction of the control group, 33 gallstone patients were divided into two subgroups, contractor group (14 cases)and non-contractor group (19 cases), The concentration of CCK-30 was significantly higher in non-contractor group than that in both contractor group and control group (55.86±3.86 pmol/l vs 37.85±0.88 pmol/l and 37.95±0.74 pmol/L, P＜0.01), but there was no difference between contractor group and control group. Meanwhile no significant difference of the concentration of CCK-f could be observed among three groups. The amount of CCK-R was lower in non-contractor group than those in both control group and contractor group (10.27±0.94 fmol/mg vs24.59±2.39 fmol/mg and 22.66±0.55 fmol/mg, P＜0.01).The activity of CCK-R shown as KD in non-contractor group decreased compared to that in control group and contractor group. Only was the activity of CCK-R lower in contractor group than that in control group. The ejection fraction correlated closely with the amount of CCK-R (r = 0.9683,P＜0.01), and the concentration of CCK-30 correlated negatively with the amount of CCK-R closely (r = -0.9627,P＜0.01).CONCLUSION: The distinctive interactive relationship of gallbladder emptying, plasma CCK and CCK-R in gallbladder from this study suggested that the defect of CCK-R may be a key point leading to the impairment of gallbladder motor function and the pathogenesis of cholesterol gallstoneformation may differ in two subgroups of gallstone patient,gallbladder non-contractor group or contractor group.

Heme oxygenase-1 overexpression increases liver injury after bile duct ligation in rats

AIM： To investigate the effects of heme oxygenase-1 （HO-1） against oxidant-induced injury caused by bile duct ligation （BDL）.METHODS： Either cobalt protoporphyrin （CoPP）, a HO-1 inducer, or saline were injected intraperitoneally in male SD-rats. Three days later, BDL or sham-operations were performed. Rats were sacrificed 3 wk after BDL and livers were harvested for histology. Fibrosis was evaluated by sirius red staining and image analysis. Alpha-smooth muscular actin, which indicates activation of stellate cells, was detected by immunohistochemical staining, and q/tokine and collagen- Iα （Col- I α） mRNA expression was detected using RNase protection assays.RESULTS： Serum alanine transaminase increased 8-fold above normal levels one day after BDL. Surprisingly, enzyme release was not reduced in rats receiving CoPP. Liver fibrosis was evaluated 3 wk after BDL and the sirius red-positive area was found to be increased to about 7.8%. However, in CoPP pretreated rats sirius redpositive areas were increased to about 11.7% after BDL. Collagen-1 α and TGF-β mRNA increased significantly by BDL. Again, this effect was increased by HO-1 overexpression.CONCLUSION： Hepatic fibrosis due to BDL is not reduced by the HO-1 inducer CoPP. In contrast, HO-1 overexpression increases liver injury in rats under conditions of experimental chronic cholestasis.

Serum insulin, insulin resistance, p-cell dysfunction, and gallstone disease among type 2 diabetics in Chinese population: A community-based study in Kinmen, Taiwan

AIM: To explore the association of serum insulin, insulin resistance, and β-cell dysfunction with gallstone disease (GSD) in type 2 diabetics. METHODS: We used a community-based study conducted between 1991 and 1993 in Kinmen, Taiwan to identify type 2 diabetics. A screening program for GSD was performed in 2001 by a panel of specialists who employed real-time ultrasound sonography to examine the abdominal region after the patient had fasted for at least 8 h. Screening was conducted in 2001 on 848 patients diagnosed with type 2 diabetes. The HOMA method was used to compare the profile differences for insulin resistance (HOMA IR) and β-cell dysfunction (HOMA β-cell). RESULTS: We studied 440 type 2 diabetics who attended sonography check-ups. After excluding eight insulin-treated diabetics, the prevalence of GSD among the remaining 432 was 13.9% (26/187) among males and 14.7% (36/245) among females. After adjustment for other GSD-associated risk factors in addition to age and obesity, GSD risk increased among females with levels of serum insulin [4th vs 1st quartile odds ratios (OR) = 4.46 (95%CI: 1.71-11.66)] and HOMA IR [4th vs 1st quartile OR = 4.46 (95%CI: 1.71-11.66)]. Better HOMA β-cell function was significantly related to decreased risk of GSD [4th vs 1st quartile OR = 0.16 (95%CI: 0.03-1.70)]. Among males, age and central obesity were the most significant risk factors for GSD. No association of GSD with serum insulin, HOMA IR, and HOMA β-cell was observed among males. CONCLUSION: Serum insulin, insulin resistance, and β-cell dysfunction are risk factors for GSD in females, but not males with type 2 diabetes.

Effects of "Plateau Bright Pearl" Green Tea on Blood Pressure,Myocardial Ultrastructure and Plasma Angiotensin Ⅱ ( Ang Ⅱ) Content in Spontaneous Hypertensive Rats

[Objective] This study aimed to further explore the relationship between tea and health. [Method] Different experimental groups were designed to preliminarily investigate the effects of ＂Plateau Bright Pearl＂ commixed green tea on blood pressure, myocardial ultrastructure and level of plasma angiotensin Ⅱ （Ang Ⅱ） in spontaneous hypertensive rats. [Result] The results showed that ＂Plateau Bright Pearl＂ commixed green tea could significantly decrease blood pressure and level of plasma angiotensin Ⅱ （ang Ⅱ） in spontaneous hypertensive rats and improve ultrastructures of left ventricular mitochondrion and myofibrillae. [Conclusion] ＂Plateau Bright Pearl＂ commixed green tea has many physiological health functions including losing weight, protecting the heart and lowering blood pressure.

Treating delayed endoscopic sphincterotomy-induced bleeding:Epinephrine injection with or without thermotherapy

AIM： To compare the hemostatic efficacy between epinephrine injection alone and epinephrine injection combined with thermotherapy for delayed postendoscopic sphincterotomy （ES） bleeding. METHODS： Cases with delayed post-ES bleeding undergoing epinephrine injection alone （epinephrine injection group, n = 26） or epinephrine combined with thermotherapy （combination therapy group, n = 33） in our institution between 1999 and 2007 were retrospectively investigated. The main outcome measurements were： initial endoscopic hemostasis, rebleeding, complications, requirement of angiographic embolization or surgery, requirement for blood transfusion, and mortality. RESULTS： The initial hemostatic efficacy was 96.2% for epinephrine injection alone and 100% for combination therapy （P = 0.44）. There were four patients with re-bleeding in each group （16.0% vs 12.1%, P = 0.72）. There was only one complication of pancreatitis from the combination therapy group. Three patients （11.5%） in the epinephrine injection group and one patient （3%） in the combination therapy group required angiographic embolization or surgery （P = 0.31）. The total number of blood transfusions was not significantly different between the two groups （3.5 ± 4.6 U vs 3.5 ± 4.5 U, P = 0.94）. There was no bleeding-related death in either group. CONCLUSION： Epinephrine injection alone is as effective as epinephrine injection combined with thermotherapy for the management of delayed post- ES bleeding.

Different distributions of interstitial cells of Cajal and platelet-derived growth factor receptor-α positive cells in colonic smooth muscle cell/interstitial cell of Cajal/plateletderived growth factor receptor-α positive cell syncytium in mice

AIM To investigate the distribution and function of interstitialcells of Cajal(ICCs) and platelet-derived growth factor receptor-α positive(PDGFRα+) cells in the proximal and distal colon.METHODS The comparison of colonic transit in the proximal and distal ends was performed by colonic migrating motor complexes(CMMCs). The tension of the colonic smooth muscle was examined by smooth muscle spontaneous contractile experiments with both ends of the smooth muscle strip tied with a silk thread. Intracellular recordings were used to assess electrical field stimulation(EFS)-induced inhibitory junction potentials(IJP) on the colonic smooth muscle. Western blot analysis was used to examine the expression levels of ICCs and PDGFRα in the colonic smooth muscle.RESULTS Treatment with NG-nitro-L-arginine methyl ester hydrochloride(L-NAME) significantly increased the CMMC frequency and spontaneous contractions, especially in the proximal colon, while treatment with MRS2500 increased only distal CMMC activity and smooth muscle contractions. Both CMMCs and spontaneous contractions were markedly inhibited by NPPB, especially in the proximal colon. Accordingly, CyPPA sharply inhibited the distal contraction of both CMMCs and spontaneous contractions. Additionally, the amplitude of stimulationinduced nitric oxide(NO)/ICC-dependent slow IJPs(sIJPs) by intracellular recordings from the smooth muscles in the proximal colon was larger than that in the distal colon, while the amplitude of electric field stimulationinduced purinergic/PDGFRα-dependent fast IJPs(fIJPs) in the distal colon was larger than that in the proximal colon. Consistently, protein expression levels of c-Kit and anoctamin-1(ANO1) in the proximal colon were much higher, while protein expression levels of PDGFRα and small conductance calcium-activated potassium channel 3(SK3) in the distal colon were much higher.CONCLUSION The ICCs are mainly distributed in the proximal colon and there are more PDGFRα+ cells are in the distal colon, which generates a pressure gradient between the two ends of the colon to propel the feces to the anus.

Cysteine peptidase and its inhibitor activity levels and vitamin E concentration in normal human serum and colorectal carcinomas

AIM: Cysteine peptidase (CP) and its inhibitor (CPI) are a matrix protease that may be associated with colorectal carcinoma invasion and progression, and vitamin E is also a stimulator of the immunological system. Our purpose was to determine the correlation between the expression of cysteine peptidases and their endogenous inhibitors,and the level of vitamin E in sera of patients with colorectal cancer in comparison with healthy individuals.METHODS: The levels of cysteine peptidases and their inhibitors were determined in the sera of patients with primary and metastatic colorectal carcinoma and healthy individuals using fluorogenic substrate, and the level of vitamin E was determined by HPLC.RESULTS: The levels of cysteine peptidases and their inhibitors were significantly higher in the metastatic colorectal cancer patients than that in the healthy controls (P＜0.05).The activity of CP increased 2.2-fold, CPI 2.8-fold and vitamin E decreased 3.4-fold in sera of patients with metastasis in comparison with controls. The level of vitamin E in healthy individuals was higher, whereas the activity of cysteine peptidases and their inhibitors associated with complexes was lower than that in patients with cancer of the digestive tract.CONCLUSION: These results suggest that the serum levels of CP and their inhibitors could be an indicator of the prognosis for patients with metastatic colorectal cancer. Vitamin E can be administered prophylactically to prevent digestive tract neoplasmas.

Development of ulcerative colitis in a patient with multiple sclerosis following treatment with interferonβ 1a

To alert clinicians to a potential novel adverse drug effect of interferonβ la, we herein report a patient with relapsing-remitting multiple sclerosis who developed ulcerative colitis following treatment with interferonβ la. Ulcerative colitis persisted despite discontinuation of interferonβ la treatment and switching the patient to glatiramer acetate. Tacrolimus （FK506）, 6-mercaptopurine, and prednisolone were required to induce remission. Both ulcerative colitis and multiple sclerosis were eventually well controlled using this regimen. Our report underscores that caution should be exercised when prescribing immunostimulatory agents in patients with inflammatory bowel disease （IBD） and challenges current efforts to stimulate innate immunity as a novel therapeutic concept for IBD.

Active chinese mistletoe lectin-55 enhances colon cancer surveillance through regulating innate and adaptive immune responses

AIM:To investigate the potential role of Active Chinese mistletoe lectin-55 (ACML-55) in tumor immune surveillance. METHODS: In this study, an experimental model was established by hypodermic inoculating the colon cancer cell line CT26 (5 × 105 cells) into BALB/c mice. The experimental treatment was orally administered with ACML-55 or PBS, followed by the inoculation of colon cancer cell line CT26. Intracellular cytokine staining was used to detect IFN-γ production by tumor antigen specific CD8+ T cells. FACS analysis was employed to profi le composition and activation of CD4+, CD8+, γδ T and NK cells. RESULTS: Our results showed, compared to PBS treated mice, ACML-55 treatment signifi cantly delayed colon cancer development in colon cancer -bearingBalb/c mice in vivo. Treatment with ACML-55 enhanced both Ag specifi c activation and proliferation of CD4+ and CD8+ T cells, and increased the number of tumor Ag specific CD8+ T cells. It was more important to increase the frequency of tumor Ag specific IFN-γ producing-CD8+ T cells. Interestingly, ACML-55 treatment also showed increased cell number of NK, and γδT cells, indicating the role of ACML-55 in activation of innate lymphocytes. CONCLUSION: Our results demonstrate that ACML-55 therapy can enhance function in immune surveillance in colon cancer-bearing mice through regulating both innate and adaptive immune responses.

Study on specific interaction of new ferrocene-substituted carborane conjugates with hemoglobin protein

The interactions between the new organometallic complexes, ferrocenesubstituted dithioocarborane conjugates （denoted as FcSB1, FcSB2 and FcSBCO） and hemoglobin （Hb） are investigated by electrochemistry, fluorescence and UVvis absorption spectroscopy. The results demonstrate that FcSB1, FcSB2 and FcSBCO can bind to the heme iron center through the replacement of the weakly bound H20/02 in the distal heme pocket of Hb by their sulfur donor atoms, inducing the allosteric change from the R state （oxygenated conformation, relax） to T state （deoxygenated conformation, tense）. The binding affinity is in the order of FcSBCO〉FeSB2〉FeSB1. Moreover, the fluorescence study illustrates that the three ferrocenecarborane conjugates differently affect the quarterly and tertiary structures as well as the polarity in the surrounding of the Trp and Tyr residues in Hb. Typically, FcSB2 mainly induces alterations of the microenvironment around the 1337Trp residue which is located on the cql32 interface of Hb. Such distinct influences are attributed to the structural features of FcSB1, FcSB2 and FcSBCO containing hydrophobic ferrocenyl and carboranyl units as well as C=O group. Screening the proteinbinding behavior can signify the potential bioactivity of such molecules and may be helpful in the future development of promising multifunctional metallodrugs.

The effect of secondary structures on the generation of characteristic events during the translocation of DNA hybrid through ?-hemolysin

Nanopore has been developed to be a powerful,single-molecule analytical tool for sensing ions,small organic molecules and biomacromolecules such as proteins and DNAs.Generally,the identity of the analyte can be revealed by current amplitude changes and mean dwell time of the analyte binding events.In some cases,generation of highly characteristic current events affords an alternative way of analyte determination with high confidence level.However,we found that secondary structures in DNA/RNA hybrids might severely hinder the generation of signature events during their translocation through?-hemolysin nanopore.In this report,we propose a strategy to add a certain concentration of urea in the buffer solution for single channel recordings and validate that low concentration of urea can effectively denature the secondary structures in DNA hybrids and recover the generation of signature events.This finding might be useful in other secondary structure-related nanopore sensing activities.

Non-surgical porto-mesenteric vein thrombosis is associated with worse long-term outcomes in inflammatory bowel diseases

Objective:Our aim was to assess the risk factors for non-surgery-related portal and mesenteric vein thrombosis(PMVT)and its impact on the outcomes of inflammatory bowel diseases(IBD).Methods:All patients with a concurrent diagnosis of IBD and PMVT between January 2004 and October 2013 were identified from the electronic medical record(study group;n=20).Patients were matched for age,sex,and IBD phenotype with control IBD patients who had no PMVT,with a ratio of 1:3(control group;n=60).Risk factors for PMVT and IBD-related outcomes at one year after diagnosis of PMVT were compared between the two groups.Results:Of the 20 patients in the Study group,6(30%)had UC,14(70%)had CD and 11(55%)were male.On multivariable analysis,inpatient status(odds ratio[OR]6.88;95%confidence interval[CI]1.88-25.12)and baseline corticosteroid use(OR 4.39;95%CI 1.27-15.19)were found to be independent risk factors for the development of PMVT.At one-year follow-up,PMVT patients were more likely to have an adverse outcome of IBD,including subsequent emergency room visit(26.3%vs.1.7%;P=0.003),hospitalization for medical management(60.0%vs.20.0%;P=0.001)or IBD-related surgery(65.0%vs.26.7%;P=0.003)than the non-PMVT controls.In multivariable analysis,PMVT(OR 5.19;95%CI 1.07-25.28)and inpatient status(OR 8.92;95%CI 1.33-59.84)were found to be independent risk factors for poor outcome,whereas baseline immunomodulator use(OR 0.07;95%CI 0.01-0.51)was found to be a protective factor.Conclusions:IBD patients who were inpatients or receiving corticosteroid therapy had an increased risk of the development of PMVT.The presence of PMVT was associated with poor clinical outcomes in IBD.

Direct evidence of the existence of specific LPS binding sites on vascular endot helial cells

Objective:To investigate the binding characteri stics of endothelial cell (EC) with LPS free from the participation of serum fac tors. Methods:Laser confocal microscope was employed in the observat ion of the binding of EC with FITC-LPS. The KD and the binding sites of each EC were calculated by radioligand binding assay of receptors (RBA) using [ 3H] -LPS. Results:The binding of EC with LPS was saturable, time and con centration dependent and it could be competed with overdosed LPS of the same typ e. The fluorescence mainly distributed in cytoplasm, especially near the nucleus , which could also be stained. Conclusions:There might be some specific LPS binding sites exi sting on ECs and LPS could function intracellularily.